Anti-Glucocorticoid Receptor alpha antibody (ab3580)
Key features and details
- Rabbit polyclonal to Glucocorticoid Receptor alpha
- Suitable for: IHC-P, ICC/IF
- Reacts with: Human
- Isotype: IgG
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Overview
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Product name
Anti-Glucocorticoid Receptor alpha antibody -
Description
Rabbit polyclonal to Glucocorticoid Receptor alpha -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
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Positive control
- IHC: human cervical carcinoma, heart tissue, tonsil tissue; ICC: U251, A2058, HeLa
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General notes
GR alpha proteins has many isoforms e.g. GR alpha-A, Alpha-2, GR-A alpha, Alpha-B. Please check Uniprot database or PMID 15866175 for more information.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.05% Sodium azide
Constituent: 99% PBS -
Concentration information loading...
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Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Assay kits
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ChIP Related Products
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Compatible Secondaries
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Conjugation kits
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Immunizing Peptide (Blocking)
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab3580 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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IHC-P | (1) |
1/20.
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ICC/IF | (1) |
1/100 - 1/200.
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Notes |
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IHC-P
1/20. |
ICC/IF
1/100 - 1/200. |
Target
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Relevance
Glucocorticoids are a family of steroids necessary for the regulation of energy metabolism and the immune and inflammatory responses. These compounds exert their effect through their interaction with the Glucocorticoid Receptor (GR) and that complex's subsequent association with DNA. All normal mammalian tissues examined to date have been shown to contain GR. The human GR exists in two forms, alpha and beta, which are thought to be the result of alternative splicing of a single gene. Sequence analysis indicates that alpha and beta forms of human GR are 777 and 742 amino acids long, respectively. They are identical up to residue 727, after which they diverge. After ligand binding, the 94 kDa GR alpha isoform translocates from the cytoplasm to the nucleus where it regulates gene expression. In contrast, the 90 kDa GR beta isoform does not appear to bind either glucocorticoid agonists or antagonists, and has been localized predominantly in the nucleus independent of hormone treatment in some human cell lines. Studies suggest that human GR alpha has a greater affinity for GR response elements (GREs) than GR beta only when in the ligand bound state. -
Cellular localization
Cytoplasm. Nucleus. Cytoplasmic in the absence of ligand; nuclear after ligand-binding. -
Database links
- Entrez Gene: 2908 Human
- SwissProt: P04150 Human
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Alternative names
- GCCR antibody
- GCR antibody
- Glucocorticoid receptor alpha isoform antibody
see all
Images
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Immunocytochemistry/Immunofluorescence analysis of U-251 MG (Human brain glioma cell line) cells labeling Glucocorticoid Receptor alpha (green) with ab3580 at 1/100. F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue). Cells were fixed with formaldehyde and incubated with the primary antibody overnight at 4°C. A DyLight 488-conjugated secondary antibody was used. 60X magnification. Right - negative control.
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Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human tonsil tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:50 with a rabbit polyclonal antibody recognizing Glucocorticoid Receptor alpha ab3580 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
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Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial adenocarcinoma cell line) cells labeling Glucocorticoid Receptor alpha (green) with ab3580 at 1/100. F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue). Cells were fixed with formaldehyde and incubated with the primary antibody overnight at 4°C. A DyLight 488-conjugated secondary antibody was used. 60X magnification. Right - negative control.
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Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human cervical carcinoma tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a rabbit polyclonal antibody recognizing Glucocorticoid Receptor alpha ab3580 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
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Immunocytochemistry/Immunofluorescence analysis of A2058 (Human metastatic melanoma cell line) cells labeling Glucocorticoid Receptor alpha (green) with ab3580 at 1/200. F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue). Cells were fixed with formaldehyde and incubated with the primary antibody overnight at 4°C. A DyLight 488-conjugated secondary antibody was used. 60X magnification. Right - negative control.
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Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human heart tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:100 with a rabbit polyclonal antibody recognizing Glucocorticoid Receptor alpha ab3580 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
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ab3580 staining glucocorticoid receptor in serum starved HeLa (Human epithelial adenocarcinoma cell line) cells treated with rosiglitazone (ab120762), by ICC/IF. Changes in localization of glucocorticoid receptor (translocation from cytoplasm to nucleous) correlates with increased concentration of rosiglitazone, as described in literature.
The cells were incubated at 37°C for 1h in media containing different concentrations of ab120762 (rosiglitazone) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab3580 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue. -
ab3580 staining Glucocorticoid Receptor alpha in mouse epididymis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with Bouin's solution and blocked with 1.5% serum for 30 minutes at 25°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/1000 in blocking buffer) for 14 hours at 4°C. ab6721 Goat anti-rabbit HRP (1/200) was used as the secondary antibody.
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ab3580 (1µg/ml) staining glucocorticoid receptor alpha in human hippocampus using an automated system (DAKO Autostainer Plus). Using this protocol there is cytoplasmic staining in the neuropil and blood vessel smooth muscle.
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
Protocols
Datasheets and documents
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Datasheet download
References (28)
ab3580 has been referenced in 28 publications.
- Lo CY et al. Increased Interleukin-17 and Glucocorticoid Receptor-β Expression in Interstitial Lung Diseases and Corticosteroid Insensitivity. Front Immunol 13:905727 (2022). PubMed: 35865549
- Vega N et al. Exposure to pollutants altered glucocorticoid signaling and clock gene expression in female mice. Evidence of tissue- and sex-specificity. Chemosphere 262:127841 (2021). PubMed: 32784060
- Ballester P et al. A transcriptional complex of NGATHA and bHLH transcription factors directs stigma development in Arabidopsis. Plant Cell 33:3645-3657 (2021). PubMed: 34586419
- Zhou Z et al. LncRNA XIST promotes inflammation by downregulating GRa expression in the adenoids of children with OSAHS. Exp Ther Med 21:500 (2021). PubMed: 33791009
- Wang SB et al. Increased lipopolysaccharide content is positively correlated with glucocorticoid receptor-beta expression in chronic rhinosinusitis with nasal polyps. Immun Inflamm Dis 8:605-614 (2020). PubMed: 32870597